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Energy Procedia 56 (2014) 112 – 117

11th Eco-Energy and Materials Science and Engineering (11th EMSES)

Synthesis of Hydroxyapatite from Oyster Shell via Precipitation

Sawittree Rujitanapanicha*, Pitoon Kumpapanb, Panthong Wanjanoic
a-c
Department of Chemistry, Faculty of Science and Technology,
Phranakhon Rajabhat University 9 Changwattana Road, Bangkhen ,Bangkok, Thailand 10220

Abstract

In this work, oyster shell which predominantly composed of calcium carbonate, was used as a staring material for synthesizing
hydroxyapatite powder. By calcinating the oyster shell powder at 1200qC for 2 h, the pure calcium oxide characterized by X-ray
diffraction (XRD), Fourier transformed infrared spectroscopy (FT-IR), was obtained. The XRD pattern also agreed with the
standard JCPDS file No.001-1160. The obtained calcium oxide powder was reacted with nitric acid to form the calcium nitrate
which was further reacted with di-ammonium hydrogen phosphate at various pH values from 8 to 10 at room temperature and
calcined at 900qC for 2 h. From XRD results, the HA powder was obtained at pH 10 as confirmed by the standard JCPDS file
No. 076-0694. In addition, The crystallize size of HA was found to be 89.5 nm from XRD result.
© 2014
© 2014Elsevier
The Authors. Published
Ltd. This is an openbyaccess
Elsevier Ltd.
article under the CC BY-NC-ND license
Peer-review under responsibility of COE of Sustainalble Energy System, Rajamangala University of Technology Thanyaburi
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
(RMUTT). under responsibility of COE of Sustainalble Energy System, Rajamangala University of Technology Thanyaburi (RMUTT)
Peer-review

Keywords: Hydroxyapatite; Oyster shell; Precipitation.

1. Introduction

Hydroxyapatite (Ca10(PO4)6(OH)2, HA) is the main mineral constituent to human bones and teeth, the major
departures in composition being a variable Ca/P mol ratio is 1.67. HA is not only a biocompatible, non-toxic, non-
immunogenic agent, non-inflammatory, but also bioactive [1]. HA can directly bond to the host bone as artificial
teeth or bones. It is widely used in orthopedic applications [2]. HA structure is hexagonal with space group P63/m,
with approximate lattice parameters a=b=9.37 Å, c=6.88 Å, D=E=90q, J=120q [3].The crystal structure of calcium

* Corresponding author. Tel.: +6-62-544-8000; fax: +6-62-522-6609.

E-mail address: sawittree_rj@yahoo.com

1876-6102 © 2014 Elsevier Ltd. This is an open access article under the CC BY-NC-ND license
(http://creativecommons.org/licenses/by-nc-nd/3.0/).
Peer-review under responsibility of COE of Sustainalble Energy System, Rajamangala University of Technology Thanyaburi (RMUTT)
doi:10.1016/j.egypro.2014.07.138
 Sawittree Rujitanapanich et al. / Energy Procedia 56 (2014) 112 – 117 113

hydroxyapatite power (Fig. 1.) synthesized by precipitation method the presence of simulated body fluid (SBF) at
37qC [2,4].
The main limitation of HA ceramic is the poor
  mechanical property. Fundamentally, all bioceramics

which have suitable for load bearing applications
PO4
should be bioinert and good mechanical properties
[5]. On the other hand, HA is one of the most
bioactive ceramics, with many medical application in
 the form of porous, dense, granules and as in coating
OH
forms [6]. HA powders can be prepared by multiple
techniques, various synthesis techniques have been
 Ca developed. These include hydrolysis of other calcium
phosphates [7], hydrothermal [2], precipitation from
aqueous solution [8], sol-gel procedure [5] and
Fig. 1. The structure of hydroxyapatite crystals. mechanochemical synthesis [9].

In the last decade, the processes involved in biomineralization has greatly developed, leading to the production of
a new generation of biomaterials.The biogenic materials such as seashells [10], animal bone [11], eggshell [12] and
Thunnus obesus bone [13]. There is a high content of calcium carbonate in oyster shell. Therefore, in the present
investigation was focused on the synthesis of HA powder from oyster shell powders via precipitation. The
crystalline phase of the powder before and after heat treatment was confirmed by XRD technique.

2. Materials and Methods

2.1 Materials
Specimens of oyster shell was collected from Surat Thani Province, Thailand. The original oyster shell was
washed in water using brush and rinsed in distilled water. After washing, oyster shell was dried in oven at 300qC
for 1 h, ground and then sieved with a 50-mesh sieve.

2.2 Synthesis of Hydroxyapatite

The oyster shell was calcined in an electrical muffle furnace at a rate of 10qC/min and maintained at various
temperatures (1000 1100 and 1200qC) for 2 h, the oyster shell transformed into calcium oxide by releasing carbon
dioxide. The products were examined by FT-IR (Shimadzu, Prestige-21) and XRD (Philips X’ Pert, USA). In the
study of synthesis HA, using the calcined that converted completely of calcium carbonate (CaCO3) to calcium oxide
(CaO). The 3.00 g of CaO obtained from oyster shell was then converted into calcium nitrate (Ca(NO3)2) in 25%
nitric acid (Merck, Germany) under constant stirring. (NH4)2HPO4 solution (12.73 g in 30 ml distilled water) was
slowly added dropwise and stirred to a solution of Ca(NO3)2 at room temperature. The reaction were maintained at
various pH value from 8 to 10, NH4OH solution was used to adjust pH of solution . The reactions that involved are
as follows:

CaCO3 CaO + CO2 (1)

CaO + 2HNO3 Ca(NO3)2 + H2O (2)

10Ca(NO3)2 + 6(NH4)2HPO4 + 8NH4OH Ca10(PO4)6(OH)2 + 20NH4NO3 + 6H2O (3)

Mixture was continued for 1 hour to achieve proper mixed solution. After completion of reaction, the solution
was subjected to aging treatment for 24 h. In the next step, the precipitated HA has been removed from solution.
The resulting HA powder was dired in air oven at 120qC for 2 h, and then calcined at 900qC for 2 h. To the best of
our knowledge, there was synthesis HA by using commercial CaO as starting reagent.
114 Sawittree Rujitanapanich et al. / Energy Procedia 56 (2014) 112 – 117

2.3 Characterization
FT-IR spectrophotometer was used to investigate the functional groups presenting in the oyster shell and pure
CaCO3. The FT-IR spectra was obtained over the region 500-4,000 cm-1 using KBr pellet technique.
The phase composition of oyster shell, calcium oxide and HA powders were studied by XRD using the Cu KD
radiation (O=0.154 nm). The data were analyzed in the 2T range from 10q to 90q with a scan step 2q per min. The
phases were identified by comparing with JCPDS. CaCO3, CaO and HA powders using the JCPDS files No. 085-
1108, 017-0912 and 076-0694 respectively.

3. Results and Discussion

3.1 Characterized of oyster shell powders

The commercial CaCO3 and original oyster shell powders were analyzed using FT-IR spectroscopy, after dried at
120qC 2 h, as shown in Fig. 2. Commercial CaCO3 bands are observed at 1496 854 and 713 cm-1 (C-O stretching)
(Fig. 2(a)). Also, bands at 1417 877 and 711 cm-1 for original oyster shell (Fig. 2(b)). Carbonate bands are also
observed at 1550-1350 cm-1, 873 cm-1 and 712 cm-1 from eggshell [12]. In Fig. 3. XRD patterns of the original
oyster shells is presented. The oyster shell patterns was identified as JCPDS (CaCO3) the file No. 085-1108. As
evident by the results, there are CaCO3 in oyster shell. As the data reported by Gil-Lim Y et al that oyster shell is
entirely composed of CaCO3 (approximately 96%) and other minerals of trivial amount [14].

(b)
Transmittance (%)

Intensity

(a)

(b) original oyster shell

(a) JCPDS (CaCO3)

4500 4000 3500 3000 2500 2000 1500 1000 500 10 20 30 40 50 60 70 80

Wavenumber (cm ) -1 2 Theta (degree)

Fig. 2. FT-IR spectra of (a) commercial calcium carbonate Fig. 3. XRD patters of (a) JCPDS 085-1108 (CaCO3)
(b) original oyster shell (b) original oyster shell

The minimum temperature for the decomposition of CaCO3 (limestone) into CaO and CO2 is 954qC, to final
5000
temperature of about 1066qC. These temperature can
4500
vary dependent upon the nature of the limestone
1200qC (e) being calcined [15]. Fig. 4. Illustrates that the XRD
4000

3500
spectra at various temperatures calcined for oyster
(d)
3000
1100qC shell powders and JCPDS (CaO) file No. 017-0912.
It can be observed that at 1000, 1100 and 1200qC
Intensity

2500
(c)
2000
1000qC
calcined, CaCO3 conversion to CaO, peak of CaO
1500 original oyster shell (b)
appears at 37.4q and 53.8q. This is consider as
1000 eggshell heated at 600-1000qC, only CaO peak exists
500
JCPDS (CaO) (a)
at 37.46q for 1000qC pattern [16]. But these crystalline
0 phase were gloomy at 1000 and 1100qC, these phases
10 20 30 40 50 60 70 80 90 100
may be attributed to the incomplete calcinations
2 Theta (degree) conditions. As a result, the best condition for
Fig. 4. XRD pattern of (a) JCPSD (CaO) file No. 017-0912 hydroxyapatite synthesis from oyster shell powders
(b) original oyster shell and original oyster shell calcined at : should be at 1200qC.
(c)1000qC; (d) 1100qC; and (e) 1200qC
 Sawittree Rujitanapanich et al. / Energy Procedia 56 (2014) 112 – 117 115

Fig. 5. shows FT-IR spectra of calcinations original oyster shell powders and commercial CaO. Result of
characteristic absorption bands for calcinations of oyster shell powders are observed at 3647 1412 and 874 cm-1
(Fig.5(a)) that were similar to commercial CaO (3642 1416 and 874 cm-1) (Fig. 5(b)). The CaO absorption band
was also reported by Namduri H., Nasrazadani S., a sharp peak at 3640 cm-1 is close to the OH band (Fig. 5(c)) [17].
 



Abs (a) (b) (c)


 
Wavenumber (cm-1) Wavenumber (cm-1) Wavenumber (cm-1)

Fig. 5. FT-IR spectra of (a) calcinations original oyster shell at temperature of 1200qC 2 h. (b) commercial CaO
(c) 100% CaO was also reported by Namduri H.

3.2 Characterized of HA-synthesis

The XRD patterns of the HA synthesized from calcinations original oyster shell powders reacted with HNO3
(NH4)2HPO4 and NH4OH solution with different pH, which was varied from 8 to 10 (Fig. 6 (e)-(g)) and powders of
commercial CaO as starting reagent (Fig. 6 (b)-(d)), are the same. The HA synthesized at pH 9 was the nearly same
as that prepared at pH 10, many sharp peaks appeared, indicating that the samples have high crystallinity. At pH 10,
the crystalline sizes at 2 theta (25-26q) of HA from CaO and oyster shell is found to be 89.5 nm. The characteristic
patterns of HA in sample closely agreed with the JCPDS file No.076-0694 (HA) (Fig. 6 (a)). In addition, the pH 8
condition was not a good condition for a crystalline HA.

* hydroxyapatite
*
* *
* * * * * * (g) pH 10

(f) pH 9

(e) pH 8
Intensity

* * ** * * * (d) pH 10

(c) pH 9

(b) pH 8

(a)JCPDS (HA)

10 20 30 40 50 60 70

2 Theta (degree)

Fig. 6. XRD patterns of HA powders synthesized and then was calcined at 900 qC for 2 h.
(a) JCPDS 076-0694 (HA) file No.076-0694; (b)-(d) synthesized from commercial CaO at pH 8 to 10;
(e)-(g) synthesized from oyster shell at pH 8 to 10.

The other method, HA powders synthesized at room temperature with buffer pH 10. The preparation of buffer pH
10, dissolve 6.8 g of ammonium chloride in 57 ml of 28-30% ammonium hydroxide and dilute 100 ml with water.
And then HA powders was dried at 120qC for 2 h (non calcined HA powders).
116 Sawittree Rujitanapanich et al. / Energy Procedia 56 (2014) 112 – 117

Fig. 7. shows the XRD patterns of HA powders synthesized from commercial CaO and oyster shell (non-
calcined). The pattern presents very broad peaks as a consequence of a poor crystallinity and crystallize size at 2
theta 25-26q is 43.5 and 44.7 nm respectively. This result agree with that reported by Martínez-Castañón G.A. et al,
diffraction patterns of the sample prepared at 25qC
for pH 10 presents very broad peaks and size of the
HA (oyster shell) crystallites is 10.12 nm [18]. HA grain gradually
increased with increasing temperature from 120 to
HA (CaO) 1200qC, and HA hexagonal-dipyramidal phase was
not transformed to the other phase up to 1200qC. At
JCPDS (HA) 1200qC, new crystalline phase are not seen, this
Intensity

precludes even amorphous phases. This result was

also the reported transformation of HA at heat-
original oyster shell treatment between 450- 1200qC by Mobasherpour et
al, calculated crystallite size from XRD pattern of
JCPDS (CaCO3) heat treatment temperature at 100 to 1200qC, with
particle in the range 7.5-59.06 nm [8], Crystal size
10 20 30 40 50 60 70 80 (DXRD) for the synthesized HA was calculated using
2 Theta (degree) the Scherrer equation: D=0.9O/EcosT, where O is the
wavelength of the X-rays (0.154 nm), T is the
Fig. 7. XRD patterns of of HA powders synthesized from scattering angle of the main reflection (25-26q), and
commercial CaO and oyster shells (non calcined HA E is the collected peak at full width at half-maximum
powders).
(FWHM) intensity [19,20].

4. Conclusion

HA powder was synthesized using oyster shell with HNO3 and (NH4)2HPO4 solution by precipitation method. The
calcinations process of original oyster shell at the temperatures of 1200qC 2 h were more complete tranformation
from CaCO3 to CaO. The prepared HA powders at different pH, results have shown that pH 10 the crystallinity was
improved. Although oyster shell is a waste material, the present investigation show that HA nanocrystalline can be
successfully prepared by a simple precipitation method at room temperature from oyster shell.

Acknowledgements

The authors are thankful to Phranakhon Rajabhat University for chemicals supporting and Asst. Prof. Dr. Apirat
Laobuthee and Mr. Attaphon Kaewvilai, Department of Materials Engineering, Faculty of Engineering, Kasetsart
University for helping and discussion.

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